Techniques - Westernblot/EUROLINE-WB

Westernblot/EUROLINE-WB

Secure differentiation of antibody results

Advantages of Westernblot/EUROLINE-WB

  • Incubation of a separate membrane strip for each serum sample.
  • The total time for performing the Western blot test is about 115 minutes.
  • All incubation steps proceed at room temperature.
  • The bands are assigned according to a lot-specific evaluation matrix.
  • A separate lot for each electrophoresis gel helps avoid errors in the assignment of the bands.
  • Every test kit contains a membrane strip of the same lot with a positive reference serum. Therefore, there is no need to incubate a positive control serum.
  • Pre-numbered membrane strips to prevent confusion.
  • Laborious labeling is not necessary.
  • Staining of the control band at the bottom of the strip showing correct completion of the individual incubation steps for each membrane strip
  • Easily and reliably differentiate positive and negative reactions. The intensity of the antigen bands correlates with the antibody titer.
  • The Western blot is the method of choice when the objective is to confirm or differentiate positive results obtained in a screening test (indirect immunofluorescence or microplate ELISA).

Test principle

  • The solid phase includes membrane strips containing electrophoretically separated antigen extracts. Therefore, the position of the proteins depends on their respective molecular masses.
  • If the sample is positive, specific antibodies in the diluted serum sample attach to the antigens coupled to the membrane.
  • In a second incubation step, the attached antibodies react with AP-labeled anti-human antibodies.
  • In a third step, the bound antibodies undergo staining with a chromogen/substrate solution which is capable of promoting a color reaction. Thus, an intense dark band at the line of the corresponding antigen appears if the serum sample contains specific antibodies.
  • Evaluating the band patterns on the incubated membrane strips involves differentiating non-specific from specific antibodies. Moreover, the number and intensity of the specific bands are decisive for the result “positive/negative”.

For product information, please contact the EUROIMMUN Sales Team

Automation

EUROBlotOne

Unique automation solution for the immunoblot work station

EUROLineScan

Automated evaluation of membrane-based test systems

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